BUGSS:Build-a-BUG:2-1

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Build-a-BUG: Series 2, Session 1

Saturday, February 9, 2013
Noon to 4:00 PM

This is the first Build-A-BUG workshop in a series of five on yeast. You will be introduced to basic lab techniques, including pipetting and centrifugation, and start your own mating type detector project. This lab will involve crash courses on molecular and cellular biology, the life cycle of budding yeast, and BioBrick-based approaches to synthetic biology, as well as hands-on minipreps (DNA isolation).

Background reading

• A Primer on Molecular Biology: Section 1.1 (pp. 1-17) provides a concise introduction for the non-biologist.
• Life Cycle of the Budding Yeast Saccharomyces cerevisiae
• Sporulation in the Budding Yeast Saccharomyces cerevisiae
• An Introduction to BioBricks
• BioBrick Assembly standard 10

Protocols

• One-Step Miniprep
• QIAGEN QuickLyse Miniprep: Read the Introduction (pp. 8-9) and Protocol (pp. 14-15)

BioBrick parts

• BBa_K801000: pTUM100 yeast shuttle vector based on pYES2
• BBa_K110006: Alpha-Cell Promoter MF(ALPHA)1
• BBa_K110005: Alpha-Cell Promoter MF(ALPHA)2
• BBa_K165057: Kozak + mCherry translational unit
• BBa_K165059: Kozak + CFPx2 translational unit
• BBa_J63002: ADH1 terminator from S. cerevisiae

Whiteboard

Yeast life cycle and sporulation

QuickLyse and One-Step miniprep protocols

BioBrick parts list

Assembly strategy (pre-cut)

Assembly strategy (post-cut)